Department of Medicine, Karolinska University Hospital and Karolinska Institutet, Solna, SE-171 76 Stockholm, Sweden.
Prostaglandins Leukot Essent Fatty Acids. 2012 Oct-Nov;87(4-5):159-63. doi: 10.1016/j.plefa.2012.07.003. Epub 2012 Aug 23.
Human 15-lipoxygenase-1 (LO) possesses mainly 15-lipoxygenase activity whereas the animal ortholog 12/15-LO possesses mainly 12-lipoxygenase activity. These findings have raised the question if studies on animals can predict the function of 15-LO-1 in human. In this study we have characterized the arachidonic acid metabolites formed by porcine 12/15-LO. Mini pigs were infected with a parasite to increase the number of blood eosinophils, which highly express 12/15-LO. Isolated porcine polymorphonuclear leukocytes (PMNL) were incubated with arachidonic acid and the produced metabolites were analysed with HPLC and mass spectrometry (MS). The cells were found to produce 15-hydroxyeicosatetraenoic acid (HETE) and 12-HETE at a ratio of 1:5. Furthermore 8,15-dihydroxyeicosatetraenoic acids (DiHETEs) and 14,15-DiHETE were formed. Based on HPLC, UV-spectroscopy and MS analysis it was found that porcine PMNL also produced eoxin (EX) C4. These results demonstrate that although porcine 12/15-LO possesses primarily 12-lipoxygenase activity, the enzyme can catalyse the formation of EXC(4).
人 15-脂氧合酶-1（LO）主要具有 15-脂氧合酶活性，而动物同源物 12/15-LO 主要具有 12-脂氧合酶活性。这些发现提出了一个问题，即在动物身上进行的研究是否可以预测人源 15-LO-1 的功能。在这项研究中，我们对猪 12/15-LO 形成的花生四烯酸代谢物进行了表征。迷你猪被寄生虫感染以增加血液嗜酸性粒细胞的数量，这些细胞高度表达 12/15-LO。用花生四烯酸孵育分离的猪嗜中性粒细胞（PMN），并用 HPLC 和质谱（MS）分析产生的代谢物。发现细胞产生 15-羟二十碳四烯酸（HETE）和 12-HETE 的比例为 1:5。此外，还形成了 8,15-二羟二十碳四烯酸（DiHETEs）和 14,15-DiHETE。通过 HPLC、UV 光谱和 MS 分析发现，猪 PMNL 还产生 eoxin（EX）C4。这些结果表明，尽管猪 12/15-LO 主要具有 12-脂氧合酶活性，但该酶可以催化 EXC（4）的形成。
